Multi-well sample testing apparatus and methods of sample testing using the same

ABSTRACT

A sample testing apparatus includes a sample tray defining a planar surface and a plurality of wells recessed relative to the planar surface, and a lid member configured to be sealed about the planar surface of the sample tray. The lid member includes an adhesive layer configured to be sealed to the planar surface of the sample tray, a breathable film layer disposed about the adhesive layer, and a backing layer disposed about the breathable film layer. Methods of using the sample testing apparatus for testing a sample and kits to facilitate such testing are also provided.

BACKGROUND

1. Technical Field

The present disclosure relates to sample testing and, more particularly,to multi-well sample testing apparatus and methods of sample testingusing the same.

2. Background of Related Art

Many industries rely on the detection and quantification of theconcentration and/or level of biological material in a liquid sample.For example, the determination of bacterial concentration in water is anessential part of water quality testing. EPA regulations require that noColiform or Escherichia coli be present in potable water. The“presence/absence” format for testing a medium is very useful in makingthis determination.

Other tests require quantification, not just detection, of bacterialconcentration in a liquid sample. Examples of such include the testingof waste water, incoming water in water purification systems, surfacewater, and food testing. Traditional methods of quantification ofbiological material include membrane filtration and the most probablenumber (MPN) method.

With respect to membrane filtration, the required volume of sample isfiltered through a membrane of a very small pore size tonon-specifically trap bacteria. The membrane is then placed on a mediumwhich supports the growth of the target bacteria. The medium isincubated at a specific temperature for a specific time and anyresulting colonies are counted.

The MPN method involves dispensing a volume of liquid sample and atesting medium into a plurality of tubes. After incubation at a specifictemperature for a specific time, the number of positive tubes iscounted. The MPN for a given volume can then be calculated based uponthe number of positive tubes, the sample volume in the negative tubes,and the total sample volume in all of the tubes. Exemplarymicrobiological quantification devices and methods using the MPN methodare detailed in U.S. Pat. Nos. 5,518,892; 5,620,895; and 5,753,456 toNaqui et al., the entire contents of each of which is incorporatedherein by reference.

SUMMARY

To the extent consistent, any of the aspects detailed herein may be usedin conjunction with any of the other aspects detailed herein.

In accordance with the present disclosure, a sample testing apparatus isprovided including a sample tray and a lid member. The sample traydefines a planar surface and includes a plurality of wells recessedrelative to the planar surface. The lid member includes an adhesivelayer configured to be sealed to the planar surface of the sample tray,a breathable film layer disposed about the adhesive layer, and a backinglayer disposed about the breathable film layer.

In aspects, the sample tray is formed from a permeable, transparent,non-toxic material. For example, the sample tray may be formed from ablend of a styrene butadiene copolymer and general purpose polystyrene.

For purposes of this application the terms “breathable” and “permeable”mean the ability to transmit gases and vapors through the barriersdisclosed herein.

In aspects, the adhesive layer is formed from a material that ispermeable, transparent, and capable of being heat sealed to the planarsurface of the sample tray. For example, the adhesive layer may beformed from ethylene vinyl acetate or modified ethylene vinyl acetate.

In aspects, the adhesive layer includes a plurality of perforations.Each perforation corresponds to and is positioned for alignment with oneof the wells defined within the sample tray such that the adhesive layerdoes not extend over any portion of the wells.

In aspects, the breathable film layer is formed from a material that ispermeable and preferably transparent. For example, the breathable filmlayer may be formed from a thermoplastic copolyester based elastomer.

In aspects, the backing layer is a paper backing layer including a paperthat is permeable, insulating, and capable of adhesive-less bonding tothe breathable film layer. For example, the backing layer may be formedfrom a kraft paper, a clay-coated paper, or an offset paper.

In aspects, the backing layer is configured as a release liner that isremovable, e.g., peelable, from the breathable film layer.

In aspects, the release liner includes a paper layer and a siliconelayer. The release liner may further include a polyester layer.

In aspects, a perforation extends through the sample tray and partiallythrough the lid member to facilitate removal of the release liner.Further, at least one connector may be provided to interconnect theportions of the sample tray on either side of the perforation. The atleast one connector may be breakable to permit removal of the releaseliner, e.g., upon initiation of peeling-off of the release liner.

In aspects, the plurality of wells include a first set of wells and asecond set of wells. In some aspects, a third set of wells is provided.Each set of wells defines a different configuration, e.g., shape,volume, etc.

In aspects, the lid member is initially sealed to the sample tray abouta first peripheral side, a second peripheral side, and a bottom end ofthe sample tray to define a pouch. An open top end of the sample tray isconfigured to permit introduction of a liquid sample into the pouch. Thelid member may further be configured to be sealed about the sample trayby heat sealing the adhesive layer to the planar surface, therebysealing each of the wells with a portion of the liquid sample therein.

In aspects, the lid member is sealed to the sample tray entirely aboutan outer perimeter of the planar surface of the sample tray to define apouch therebetween. In such aspects, the sample tray defines a slitextending therethrough that is configured to permit introduction of aliquid sample into the pouch.

In aspects, the sample tray includes at least one reinforcement memberconfigured to provide structural support to the sample tray.

In aspects, the sample tray defines at least one bridge fluidlyconnecting adjacent wells to facilitate the capture of an air bubblewithin at least one of the adjacent wells upon sealing of the lid memberwith the sample tray.

Another sample testing apparatus provided in accordance with the presentdisclosure includes a sample tray and a lid member. The sample traydefines a planar surface and includes a plurality of wells recessedrelative to the planar surface. The lid member is configured to besealed to the planar surface of the sample tray to seal each of theplurality of wells. An insert positioned between the sample tray and thelid member is configured to capture an air bubble within at least one ofthe wells.

In aspects, the insert includes a dissolvable film configured todissolve upon contact with a liquid sample. The dissolvable film definesat least one well. The at least one well of the dissolvable film isconfigured for positioning within a corresponding well of the sampletray. More specifically, the at least one well of the dissolvable filmmay define a reduced depth relative to the corresponding well of thesample tray to define an air pocket therebetween. Further, thedissolvable film may be formed from polyvinyl alcohol.

In aspects, the insert includes a plate defining a plurality ofcut-outs. Each cut-out is configured for positioning about acorresponding well and defines a reduced opening in at least onedimension as compared to an opening of the corresponding well.

Another sample testing apparatus provided in accordance with the presentdisclosure includes a sample tray and a lid member. The sample traydefines a planar surface and includes a plurality of wells recessedrelative to the planar surface. The lid member is configured to besealed to the planar surface of the sample tray to seal each of theplurality of wells. At least one bridge is provided to fluidly connectadjacent wells. The at least one bridge is configured to facilitate thecapture of an air bubble within at least one of the adjacent wells uponsealing of the lid member with the sample tray.

Another sample testing apparatus provided in accordance with the presentdisclosure includes a sample tray and a lid member. The sample traydefines a planar surface and includes a plurality of wells recessedrelative to the planar surface. The lid member is configured to besealed to the planar surface of the sample tray to seal each of theplurality of wells. At least one dissolvable capsule is provided. Eachdissolvable capsule is disposed within one of the wells of the sampletray and is configured to dissolve to provide an air bubble within thewell.

A method of testing a sample provided in accordance with the presentdisclosure includes providing a sample testing apparatus including asample tray and a lid member. The sample tray defines a plurality ofwells including a first set of wells having a first configuration and asecond set of wells having a second configuration. Additional sets,e.g., a third set of wells having a third configuration, are alsocontemplated. The lid member is sealed to the sample tray about at leasta first peripheral side, a second peripheral side, and a bottom end ofthe sample tray and defines a pouch therebetween. The method furtherincludes introducing a predetermined volume of a liquid sample into thepouch of the sample testing apparatus and sealing the lid member to thesample tray to seal each of the plurality of wells such that each of thewells of the first set of wells is filled to capacity with a firstportion of the liquid sample and such that a remainder of the liquidsample is evenly distributed into the wells of the second set of wells.The volume of liquid sample in each of the wells of the second set ofwells may be less than a volume capacity of each of the wells in thesecond set of wells.

In aspects, sealing the lid member to the sample tray is performed viaheat sealing. The sample tray may be positioned within a receptacleprior to heat sealing. Further, the sample tray may be enclosed withinthe receptacle using a cover flap prior to heat sealing.

In aspects, the method further includes incubating the sealed sampletesting apparatus. Incubating the sealed sample testing apparatus may beperformed with the lid member facing downward, i.e., inverted.

In aspects, the method further includes peeling off a backing layer ofthe lid member prior to incubating the sealed sample testing apparatus.

In aspects, peeling off the backing layer includes grasping the backinglayer at a central apex thereof and peeling off the backing layer suchthat the peeling off is initiated adjacent a center of the sample tray.Alternatively, peeling may be initiated from either top corner of thesample tray.

In aspects, the method further includes counting a number of positivewells and determining a result based upon the number of positive wells.

In aspects, the lid member is sealed to a bottom of the sample tray andunsealed at a top of the sample tray. In such aspects, introducing thepredetermined volume of the liquid sample into the pouch includesintroducing the predetermined volume through the unsealed top.

In aspects, the lid member is sealed about a perimeter edge of thesample tray. In such aspects, introducing the predetermined volume ofthe liquid sample into the pouch includes introducing the predeterminedvolume through a slit defined within the sample tray.

A kit provided in accordance with the present disclosure includes asample testing apparatus and a receptacle. The sample testing apparatusgenerally includes a sample tray defining a plurality of wells, and alid member disposed to cover the plurality of wells. The sample testingapparatus may further be configured to include any of the aspectsdetailed above. The receptacle is configured to receive the sampletesting apparatus and includes a base portion and, in some aspects, acover flap. The base portion defines a cavity for receipt of the sampletesting apparatus therein. The cover flap is releasably engagable withthe base portion. When engaged to the base portion, the cover flap ispivotable relative to the base portion between an open positionpermitting insertion and withdrawal of the sample testing apparatus fromthe cavity, and a closed position wherein the cover flap encloses thesample testing apparatus within the receptacle.

In aspects, the base portion includes a cut-out defined adjacent anouter peripheral edge thereof. The cut-out is configured to facilitateinsertion or withdrawal of the sample testing apparatus to/from thecavity.

In aspects, the cavity includes a plurality of discrete chambers. Eachchamber is configured to receive one of the plurality of wells of thesample testing apparatus upon insertion of the sample testing apparatusinto the cavity.

In aspects, the base portion of the receptacle defines an engagementslot and the cover flap includes an engagement pin coupled thereto. Theengagement pin is configured for snap-fit engagement within theengagement slot to releasably couple the cover flap to the base portion.More specifically, the engagement slot may include a shoulder configuredto inhibit withdrawal of the engagement pin from the engagement slot, orthe engagement slot may include a neck portion and an enlarged portionwherein the neck portion inhibits withdrawal of the engagement pin fromthe enlarged portion.

In aspects, the base portion of the receptacle includes at least onefinger spaced-apart from the base portion to define an engagement areatherebetween. In such aspects, the cover flap includes an engagement pincoupled thereto that is configured for releasable positioning within theengagement area to releasably couple the cover flap to the base portion.

In aspects, the base portion of the receptacle defines at least onefirst lumen and the cover flap defines at least one second lumen. Thefirst and second lumens are configured to align with one another topermit insertion of an engagement pin therethrough to releasably couplethe cover flap to the base portion.

BRIEF DESCRIPTION OF THE DRAWINGS

Various aspects of the present disclosure are described herein withreference to the drawings wherein like reference numerals identifysimilar or identical elements:

FIG. 1A is an exploded, perspective view of a multi-well sample testingapparatus provided in accordance with the present disclosure;

FIG. 1B is an exploded, perspective view of another multi-well sampletesting apparatus provided in accordance with the present disclosure;

FIG. 2 is a bottom, perspective view of the apparatus of FIG. 1A;

FIG. 3A is a cross-sectional view of the apparatus of FIG. 1A takenalong section line “3A-3A” of FIG. 2;

FIG. 3B is a cross-sectional view of the apparatus of FIG. 1A takenalong section line “3B-3B” of FIG. 2;

FIG. 4A is an enlarged, perspective, cut-away view of a sample tray ofanother multi-well sample testing apparatus, including a water-solublefilm disposed thereon;

FIG. 4B is an enlarged, perspective, cut-away view of a sample tray ofanother multi-well sample testing apparatus, including capsules disposedwithin the wells thereof;

FIG. 5A is an enlarged, perspective, cut-away view of a sample tray ofanother multi-well sample testing apparatus, including a bridgeconnecting adjacent wells to one another;

FIG. 5B is an enlarged, perspective, cut-away view of a sample tray ofanother multi-well sample testing apparatus, including another bridgeconnecting adjacent wells to one another;

FIG. 6A is an exploded, perspective view of a sample tray of anothermulti-well sample testing apparatus, including a mask disposed thereon;

FIG. 6B is an exploded, perspective view of the sample tray and mask ofFIG. 6A including a lid member disposed thereon;

FIG. 7A is a bottom, perspective view of another multi-well sampletesting apparatus provided in accordance with the present disclosure anddisposed in a closed condition;

FIG. 7B is a partial, bottom, perspective view of a the apparatus ofFIG. 7A, disposed in an open condition;

FIG. 8 is a bottom, perspective view of another multi-well sampletesting apparatus provided in accordance with the present disclosure;

FIG. 9 is a bottom, perspective view of another multi-well sampletesting apparatus provided in accordance with the present disclosure;

FIG. 10A is a bottom, perspective view of another multi-well sampletesting apparatus provided in accordance with the present disclosure;

FIG. 10B is a side view of the apparatus of FIG. 10A;

FIG. 10C is an enlarged view of the area of detail indicated as “10C” inFIG. 10B;

FIG. 11 is a transverse, cross-sectional view of a portion of anotherlid member provided in accordance with the present disclosure andincluding a release liner;

FIG. 12 is a transverse, cross-sectional view of a portion of anotherlid member provided in accordance with the present disclosure andincluding a release liner;

FIG. 13A is a perspective view of a sealing insert provided inaccordance with the present disclosure, shown including a multi-wellsample testing apparatus positioned therein;

FIG. 13B is a perspective view of a cover of the sealing insert of FIG.13A;

FIG. 13C is a transverse, cross-sectional view of the engagement betweenthe cover and a base of the sealing insert of FIG. 13A;

FIG. 14 is a perspective view of another sealing insert provided inaccordance with the present disclosure;

FIG. 15 is a perspective view of another sealing insert provided inaccordance with the present disclosure;

FIG. 16A is a perspective view of another sealing insert provided inaccordance with the present disclosure;

FIG. 16B is a perspective view of a base of the sealing insert of FIG.16A;

FIG. 16C is an enlarged view of the area of detail indicated as “16C” inFIG. 16B; and

FIG. 16D is an enlarged perspective view of the engagement end of thecover of the sealing insert of FIG. 16A.

DETAILED DESCRIPTION

Provided in accordance with the present disclosure and detailed beloware apparatus and methods that facilitate the detection and/orquantification of biological material, e.g., bacteria, fungi or otherliving organisms, aggregates of proteins such as enzymes, co-factorsusing reaction mixtures, etc., within a liquid sample. A testing medium,e.g., chemical and/or microbiological reactants, suitable for enablingdetection of the specific biological material to be quantified isintroduced into the liquid sample prior to testing. As can beappreciated, the testing medium utilized will depend on the biologicalmaterial to be detected. More specifically, a testing medium is selectedthat enables detection of the presence of the biological material soughtto be quantified, preferably does not detect the presence of otherbiological material likely to be present in the liquid sample, andprovides a sensible change, e.g., color change, fluorescence, etc., ifthe biological material sought to be detected is present in the liquidsample. Exemplary tests capable of being performed using the apparatusand methods of the present disclosure include the detection of:Coliforms and E. coli, Legionella, Enterococci, and Pseudomonasaeruginosa (e.g., using the Pseudalert® test kit, manufactured by IDEXXLaboratories, Inc. of Westbrook, Me., USA). Other suitable tests arealso contemplated.

Turning to FIGS. 1A-1B, multi-well sample testing apparatus 100, 200provided in accordance with the present disclosure are shown eachgenerally including a lid member 110, 210 and a sample tray 140, 240.The lid members 110, 210 and sample trays 140, 240 of the respectiveapparatus 100, 200 define generally rectangular configurations ofsimilar dimension and are sealingly engaged to one another along threeedges of the apparatus 100, 200, e.g., side edges 102, 104 and 202, 204and bottom edges 106, 206, respectively, so as to define a pouch-likeconfiguration. Open top edges 108, 208 of the apparatus 100, 200 permitthe introduction of the liquid sample to be tested (including thetesting medium) into the interior of the pouch, as will be detailedbelow.

With reference to FIG. 1A, the lid member 110 of the apparatus 100 isformed from three layers of material: an adhesive layer 112, abreathable film layer 114, and a backing layer 116. The backing layer116 is optional and preferably paper. The adhesive layer 112 isconfigured to interface directly with the sample tray 140 to adhere thelid member 110 to the sample tray 140 upon heat sealing, as will bedetailed below. The material forming the adhesive layer 112 is selectedbased upon its ability to seal to the sample tray 140 via heat sealing,provide some degree of permeability without allowing for excess liquidsample loss during incubation, and for its transparency. In embodiments,the adhesive layer 112 is formed from a relatively thin sheet ofethylene vinyl acetate or modified ethylene vinyl acetate.

The breathable film layer 114 of the lid member 110 is disposed betweenthe adhesive layer 112 and the paper backing layer 116. The materialforming the breathable film layer 114 is selected based upon itspermeability, ability to withstand the relatively high temperatures ofheat sealing, ability to withstand and not discolor duringsterilization, transparency, and optimization with testing media, e.g.,indicator reagents. In embodiments, the breathable film layer 114 isformed from a thermoplastic copolyester based elastomer. In otherembodiments, the breathable film layer 114 may be formed from ethylenevinyl acetate or modified ethylene vinyl acetate, a thermoplasticcopolymer, a thermoplastic polyurethane elastomer or aromatic polyether,a styrene butadiene copolymer, or fluorinated ethylene propylene.

The paper backing layer 116 of the lid member 110 is disposed on thebreathable film layer 114. The material forming the paper backing layer116 is selected based upon its permeability, ability to bond with thebreathable film layer 114 without the need for an adhesive therebetween,ability to withstand the relatively high temperatures of heat sealing,printability, and insulating properties (i.e., the ability to allow theproper amount of heat transfer to the adhesive layer 112 during heatsealing). In embodiments, the paper backing layer 116 is formed from akraft paper, or may alternatively be formed from an offset paper(preferably of 50 lb. weight, although other weights are alsocontemplated). Further, the paper backing layer 116 may be colored whiteto provide a suitable background to facilitate detection of a colorchange of the testing medium. Alternatively, the paper back layer 116may be removable, e.g., peelable, after heat sealing of the lid member110 to the sample tray 140 to expose the transparent breathable filmlayer 114. In such configurations, rather than providing a whitebackground, apparatus 100 may be positioned adjacent a different colorbackground or a light box to facilitate detection of a change in thetesting medium. Providing a removable paper back layer 116 alsoincreases the permeability of the lid member 110 during incubation byexposing the breathable film layer 114 and inhibits liquid sample lossduring incubation due to the paper back layer 116 acting as a desiccant.The paper backing layer 116 may further include a tab 117 extendingtherefrom adjacent the open top edge 108 of the apparatus 100 tofacilitate opening and/or handling of apparatus 100. In someembodiments, paper backing layer 116 is omitted.

With respect to manufacturing the lid member 110, the breathable filmlayer 114 is extruded and cast directly onto the paper backing layer116, and the adhesive layer 112 is extruded and cast directly onto thebreathable film layer 114. Alternatively, the adhesive layer 112 and thebreathable film layer 114 may be cast onto the paper backing layer 116during co-extrusion of the adhesive and breathable film layers 112, 114,respectively. The layers 112, 114, 116 may also be formed as separatefilms and laminated together.

Referring to FIG. 1B, the apparatus 200 is similar to the apparatus 100(FIG. 1A) except for the configuration of the lid member 210. Theapparatus 200 may further include any or all of the features of theapparatus 100 (FIG. 1A). For purposes of brevity, only the differencesbetween the apparatus 200 and the apparatus 100 (FIG. 1A) are detailedbelow.

The lid member 210 of the apparatus 200 is formed from at least threelayers of material: an adhesive layer 212, a breathable film layer 214,and a paper backing layer 216. The breathable film layer 214 and thepaper backing layer 216 are similar to those detailed above with respectto the lid member 110 (FIG. 1A). The adhesive layer 212 differs from theadhesive layer 112 of the lid member 110 (FIG. 1A) in that the adhesivelayer 212 is perforated, e.g., via die cutting, laser cutting, or othersuitable manufacturing process, to remove the portions disposed over thewells 244 of the sample tray 240. As a result, during sealing, theadhesive layer 212 is sealed to the sample tray 240 over the entiresurface 242 thereof, between and about each of the wells 244 definedwithin the sample tray 240, but does not extend across any of the wells244 defined within the sample tray 240. This configuration providesincreased permeability without compromising sealing integrity.

Referring again to FIG. 1A, and with additional reference to FIGS. 2-3B,the sample tray 140 of the apparatus 100 defines a generally planarupper surface 142 and includes a plurality of sets of wells 144, 146,148 of varying configuration recessed relative to the generally planarupper surface 142. More specifically, the sample tray 140 includes aplurality of small wells 144 positioned towards the sealed bottom edge106 of the apparatus 100, a plurality of medium wells 146 positionedintermediate the sets of wells 144, 148, and a plurality of elongatedlarge wells 148 positioned towards the open top edge 108 of theapparatus 100. The different configurations of the wells 144, 146, 148eliminate the need to dilute the liquid sample and increase the range ofquantification. Each of the sets of wells 144, 146, 148 will bedescribed in greater detail, in turn, below. However, otherconfigurations of wells of similar and/or varying configuration are alsocontemplated such as, for example, those disclosed in the '892, '895,and '456 patents to Naqui et al., previously incorporated herein byreference.

The material forming the sample tray 140 is selected based upon itspermeability, non-toxicity to the biological material being detected,ability to withstand and not discolor during sterilization,transparency, reduced or absent fluorescence, and its ability to besealed to the adhesive layer 112 via heat sealing. In embodiments, thesample tray is formed from a blend of a styrene butadiene copolymer andgeneral purpose polystyrene. This blend has been found to achieve theabove-noted criteria. In particular, this blend has been found to beparticularly suited for sealing to ethylene vinyl acetate or modifiedethylene vinyl acetate, the material used to form the adhesive layer112. Further, it has been found that this blend is advantageous in thatit is non-toxic with respect to many biological materials to bedetected, e.g., Legionella bacteria, while polyvinyl chloride has beenfound to be toxic to and kill certain biological materials, e.g.,Legionella bacteria.

Referring to FIGS. 2-3B, the plurality of small wells 144 are arrangedin a three by ten matrix positioned towards the sealed bottom edge 106of apparatus 100. Each small well 144 is generally rectangular inconfiguration, although the side walls of each small well 144 angleslightly inwardly towards one another in the direction approaching thebase of the small well 144. The relatively shallow configuration of thesmall wells 144 maximizes the ratio of lid member surface area above thewell 144 to liquid sample volume retained within the well 144. It hasbeen found that maximizing this ratio within sample wells helps optimizebacteria growth and maximize permeability. Preferably, this ratio isequal to or greater than about 0.33 cm²/mL for all wells. Inembodiments, each small well 144 defines a volume of 0.20 mL, isconfigured to receive about 0.20 mL of liquid sample (100% capacity),and defines a lid member surface area (in cm²) to sample volume (in mL)ratio of about 0.67.

The plurality of medium wells 146 are arranged in a five by six matrixpositioned between the small wells 144 and the large elongated wells148. Each medium well 146 defines an inverted pyramid configuration witha rounded or flattened base. This configuration provides each of themedium wells 146 with an increased depth without requiring an increasein volume. This increased depth provides a longer “view path” throughthe liquid sample in the well 146. The longer “view path” providesbetter color differentiation, thus facilitating detection of whetherthere is a color change of the liquid sample (due to the testing medium)within the well 146. The inverted pyramid configuration of the mediumwells 146 also maximizes the ratio of lid member surface area above thewell 146 to liquid sample volume retained within the well 146. Inembodiments, each medium well 146 defines a volume of about 1.01 mL, isconfigured to receive about 1.01 mL of liquid sample (100% capacity),and defines a lid member surface area (in cm²) to sample volume (in mL)ratio of about 0.48.

The plurality of elongated large wells 148, e.g., six elongated largewells 148, are arranged to extend longitudinally in side-by-siderelation and are positioned towards the open top edge 108 of apparatus100. With the small and medium wells 144, 146, respectively, filled withthe liquid sample to 100% capacity, the large elongated wells 148receive the remainder of the liquid sample. The elongated configurationof the large wells 148 and the relatively shallow depth of the largewells 148 enables the large wells 148 to retain a relatively largevolume of liquid sample without compromising the ratio of lid membersurface area above the wells 148 to liquid sample volume retained withinthe wells 148. In embodiments, each large elongated well 148 defines avolume of about 18.86 mL, is configured to receive about 10.95 mL ofliquid sample (about 58% capacity, with the remaining volume occupied byair), and defines a lid member surface area (in cm²) to sample volume(in mL) in each well 148 ratio of about 0.35. The about 58% capacity ofthe large elongated wells 148, as will be detailed below, is a result ofthe liquid sample first being utilized to fill the small and mediumwells 144, 146, respectively, to capacity using a 100 mL sample, withthe remaining liquid sample then being equally distributed into thelarge wells 148. However, as also detailed below, in other embodiments,various features may be provided to capture air bubbles within the wellsto achieve an appropriate percentage of capacity occupied by the liquidsample.

Other suitable numbers, arrangements, and/or configurations of the wells144, 146, 148 of the apparatus 100 are also contemplated, depending on aparticular purpose. Further, a reduced thickness at the base of any orall of the wells 144, 146, 148 (or the bases of any or all of the wellsof any other suitable sample tray) may be provided to increasepermeability without compromising structural stability of the wells 144,146, 148.

It has been found that capturing air or air bubbles within some or allof the wells of a sample tray helps optimize bacterial growth withrespect to many bacteria to be detected. In particular, the percentageby volume of liquid sample within each well (or some of the wells) maybe in the range of about 50% to about 65% (about 50% to about 65%capacity), while the remaining about 35% to about 50% percentage byvolume is occupied by air and/or air bubbles. Features which may beincorporated into the wells to achieve air bubbles in this or anothersuitable percentage by volume range include: configuring the wells todefine diamond, tear drop, or hour-glass configurations, and/orincluding indentations, sharp corners, protrusions, or other geometricfeatures within any or all of the wells. Other additional or alternativefeatures for this purpose and configured for use with the apparatus 100,200 (FIG. 1B), or any other suitable sample testing apparatus, aredetailed below with respect to FIGS. 4-5B.

Turning to FIG. 4A, as noted above, capturing an air bubble within thesample wells has been found to help optimize bacterial growth.Accordingly, a dissolvable film 350 may be provided for positioningbetween the sample tray 340 and the lid member (not shown) of theapparatus 300. The apparatus 300 may be similar to the apparatus 100(FIG. 1A) or the apparatus 200 (FIG. 1B), and may include any of thefeatures thereof, but differs at least in that the apparatus 300 furtherincludes the dissolvable film 350 disposed between the sample tray 340and the lid member (not shown) thereof.

The dissolvable film 350 is dimensioned and configured similar to thesample tray 340 except that the wells 352 defined within the dissolvablefilm 350 define reduced depths as compared to the corresponding wells344 of the sample tray 340. As a result of this configuration, an airpocket 355 is defined between the bases of the wells 352 of thedissolvable film 350 and the bases of the corresponding wells 344 of thesample tray 340. The dissolvable film 350 is configured to dissolve uponcontact with the liquid sample without hindering the biological materialto be detected or the testing medium contained within the liquid sample.As the liquid sample enters each well 344 and the dissolvable film 350dissolves, the air disposed within the air pocket 355 is captured in theform of an air bubble within each well 344. In embodiments, thedissolvable film 350 is a water soluble film formed from polyvinylalcohol.

Referring to FIG. 4B, another embodiment of a sample testing apparatus3000 is provided including a sample tray 3400 and lid member (notshown). Sample testing apparatus 3000 may be similar to apparatus 100,200 (FIGS. 1A and 1B, respectively), or any of the other embodimentsdetailed herein, and may include any of the features thereof. Sampletesting apparatus 3000 differs from the other embodiments detailedherein, or additionally includes, as detailed below, a dissolvablecapsule 3500 disposed within one or more of wells 3450 in order tocapture an air (or other gas or gas mixture) bubble within the samplewell(s) 3450 to facilitate bacterial growth.

Each dissolvable capsule 3500 is configured for positioning within oneof wells 3450 of sample tray 3400 and may be placed therein or adhered,e.g., using resin, to an interior surface thereof, e.g., base 3460 orany of sidewalls 3470. Capsules 3500 may be positioned and/or adheredwithin their respective wells 3450 during manufacturing of sample tray3400, or may be positioned therein at the user-end. Each dissolvablecapsule 3500 is formed from a dissolvable material. In embodiments,capsules 3500 are water soluble capsules such as hydroxypropylmethylcellulose (HPMC) capsules, gelatin capsules, or other suitablewater soluble capsules capable of remaining substantially intact duringthe heat sealing process and, ultimately, sufficiently dissolving duringincubation of sample testing apparatus 3000 to thereby create an air (orother gas or gas mixture) bubble within the sealed well 3450.

Each dissolvable capsule 3500 is formed from two capsule portions 3510,3520, one of which is inserted partially into the other to define anoverlapping region 3530 and an enclosed interior chamber 3540.Dissolvable capsules 3500 may be assembled from capsule portions 3510,3520 in a normal environment to entrap air within interior chamber 3540,or may be assembled in a special environment, e.g., an oxygen-richenvironment, to entrap a desired gas composition, e.g., oxygen-enrichedair, within interior chamber 3540. Further, dissolvable capsules 3500are configured so as to not interfere with the biological material to bedetected or the testing medium contained within the liquid sample.

As noted above, dissolvable capsules 3500 are configured to bedissolved, e.g., via the liquid sample, to create an air (or other gasor gas mixture) bubble within the sealed well 3450. More specifically,capsules 3500 are configured, e.g., the thickness of and/or materialforming capsules are selected, such that capsules 3500 begin to dissolveupon contact with the liquid sample but at a sufficiently slow rate soas to maintain the interior chamber 3540 in a sealed condition duringheat sealing. Ultimately, during incubation, capsules 3500 aresufficiently dissolved so as to enable the escape of the air and/orgas(es) from interior chamber 3540 into the sealed well 3450 to providean air (or other gas or gas mixture) bubble therein.

With reference to FIGS. 5A-5B, as shown with respect to apparatus 400,500, air bubbles may also be captured by providing relatively shallowbridges interconnecting adjacent wells of the sample tray. For example,as shown in FIG. 5A, the sample tray 440 may include linear bridges 443interconnecting adjacent wells 444 to one another in any suitablepattern or configuration. Alternatively, as shown in FIG. 5B, the sampletray 540 may include curved, or “U”-shaped bridges 543 interconnectingadjacent wells 544 to one another in any suitable pattern orconfiguration. Other configurations of bridges are also contemplated. Ineither configuration, the bridges 443, 543, which define reduced depthsas compared to the wells 444, 544, respectively, facilitate capturingair bubbles in the wells 444, 544 during sealing. Alternatively, a maskcan be used to fully enclose one of the two interconnected wells,thereby trapping air in the enclosed well, leaving the partner well opento fill with fluid.

Referring to FIGS. 6A-6B, as shown with respect to apparatus 600, airbubbles may also be captured by controlling the flow of the liquidsample into the wells using a mask plate 660 positioned between the lidmember 610 and the sample tray 640. The mask plate 660 is dimensionedsimilar to the generally planar upper surface 642 of the sample tray 640and includes a plurality of cut-outs 662, each of which corresponds toone of the wells 644 formed within the sample tray 640. However,although the cut-outs 662 are generally aligned with their correspondingwells 644, the cut-outs 662 preferably define a reduced opening in atleast one dimension, e.g., the length and/or width dimension, ascompared to the opening of the corresponding wells 644. Thisconfiguration creates a bottleneck effect and enables the capture of airbubbles within each of the wells 644 as the liquid sample flows throughthe reduced-dimension cut-outs 662 and into the relativelylarger-dimensioned wells 644.

Referring generally to FIGS. 1A and 2-3B, the use of apparatus 100 forquantifying a bacterial concentration in a liquid sample is detailed. Asnoted above, apparatus 200, 300, 400, 500, 600 (FIGS. 1A, 4, 5A, 5B, and6A-6B, respectively) are similar to and may include any of the featuresof apparatus 100, and vice versa. Accordingly, the use thereof issimilar to that of apparatus 100 and will not be detailed hereinbelowfor purposes of brevity.

Initially, a suitable testing medium, selected based upon the biologicalmaterial to be detected, is introduced into the liquid sample and 100 mLof the liquid sample, including the testing medium, is measured. Themeasured 100 mL of liquid sample is then introduced, e.g., poured, intothe pouch of the apparatus 100 through the open end 108 thereof. Tofacilitate the introduction of the liquid sample, the side edges 102,104 of the apparatus 100 may be squeezed towards one another to slightlybend the lid member 110 and/or the sample tray 140 to enlarge the pouchopening defined between the lid member 110 and the sample tray 140.

With the liquid sample disposed within the pouch between the lid member110 and the sample tray 140, the apparatus 100, lead by open end 108,may be fed into a heat sealer (e.g., a heat sealer sold under the nameQuanti-Tray® Sealer 2X, manufactured by IDEXX Laboratories, Inc. ofWestbrook, Me., USA). As the apparatus 100 is translated through theheat sealer, the heat sealer urges the lid member 110 into contact withthe sample tray 140 such that the liquid sample is first evenlydistributed into the small wells 144 at a full capacity volume of 0.20mL, is then evenly distributed into the medium wells 146 at a fullcapacity volume of 1.01 mL, and the remaining liquid is evenlydistributed into the large wells 148 (about 10.95 mL of liquid sample ineach of the large wells 148 filling these large wells to about 58%capacity (or between about 50% and about 65% capacity, depending on aparticular purpose). The air filling the remainder of these large wells148 after sealing facilitates bacterial growth in these wells 148,similarly as detailed above with respect to the capture of air bubbles.

Simultaneously with or near-simultaneously with the urging of the lidmember 110 and the sample tray 140 into contact with one another todistribute the liquid sample, heat applied to the apparatus 100 via theheat sealer effects heat-sealing of the adhesive layer 112 to the sampletray 140 fully about the surface 142 of the sample tray 140 to sealinglyenclose each of the wells 144, 146, 148. The insulative properties ofthe paper backing layer 116 of the lid member 110 enable heat sealing ofthe adhesive layer 112 with the sample tray 140 but inhibit the liquidsample from being significant effected, i.e., heat sealing is effectedwith minimal temperature increase of the liquid sample.

Once the apparatus 100 has been sealed, it is incubated for apre-determined amount of time under pre-determined conditions (dependingupon the test being performed). In embodiments, the apparatus 100 (orother suitable apparatus) is incubated in an inverted orientation. Thisconfiguration allows the liquid sample within each of the wells 144,146, 148 to directly contact and sit atop the lid member 110, while anyair trapped in the wells, e.g., large wells 148, is positioned betweenthe liquid sample and the base of the well 148. This configuration hasbeen found to contribute to better bacteria growth. However, otherincubation orientations are also contemplated.

After the incubation period, the results are determined, recorded, andanalyzed. In embodiments where the paper backing layer 116 of the lidmember 110 is removable, the paper backing layer 116 may be removedprior to incubation, to increase permeability, or after incubation, tofacilitate determining the results. In order to determine the quantityof the biological material being tested for in the liquid sample, thenumber of “positive” wells are counted, as indicated by a color changeor other sensible change in the well, and routine statistical analysisis performed (or a look-up table including pre-calculated statisticalresults of such is utilized).

Turning now to FIGS. 7A-10C, various additional embodiments ofmulti-well sample testing apparatus provided in accordance with thepresent disclosure are shown and described. Although shown as separateembodiments to highlight particular features of each of the apparatus,it is contemplated that any of features of the below embodiments beutilized in conjunction with one another and/or any of the features ofthe above embodiments, except where specifically contradicted.

Referring to FIGS. 7A and 7B, multi-well sample testing apparatus 700generally includes a lid member 710 and a sample tray 740. The lidmember 710 and sample tray 740 define generally rectangularconfigurations of similar dimensions, except that the lid member 710 andsample tray 740 both define similar outwardly-bowed top portions 717,747, respectively. The outwardly-bowed top portions 717, 747 facilitatethe grasping and manipulation of the apparatus 700 and, as detailedbelow, the opening of apparatus 700 to permit the introduction of theliquid sample to be tested therein.

The lid member 710 and sample tray 740 are sealingly engaged to oneanother along all four outer peripheral edges of the apparatus 700,e.g., side edges 702, 704, bottom edge 706, and top edge 708, such thatthe outer periphery of the apparatus 700 is fully sealed. This is incontrast to apparatus 100, 200 (FIGS. 1A and 1B, respectively), detailedabove, wherein only three of the edges are sealed. Rather than providingan open edge to permit introduction of the liquid sample into theinterior of the apparatus 700, the apparatus 700 includes a slit 780defined within and extending through the planar upper surface 742 of theoutwardly-bowed top portion 747 of the sample tray 740, as will bedetailed below. The lid member 710 may otherwise be configured similarlyas any of the other embodiments detailed herein and may include any ofthe features and/or combination of features thereof.

The sample tray 740 of the apparatus 700 defines a generally planarupper surface 742 and includes two sets of wells 744, 748 of differentconfigurations recessed relative to the generally planar upper surface742. More specifically, the sample tray 740 includes a plurality ofsmall wells 744, e.g., a 9×10 matrix of small wells 744, positionedtowards the bottom edge 706 of the apparatus 700, and a plurality ofelongated large wells 748, e.g., six elongated large wells 748,extending longitudinally between the small wells 744 and outwardly-bowedtop portion 747 of the sample tray 740. The small wells 744 areconfigured to be filled with the liquid sample to 100% capacity with theremainder of the sample being distributed into (but not filling) theelongated large wells 748. Other configurations of wells of similarand/or varying configuration are also contemplated, such as thosedetailed above.

Continuing with reference to FIGS. 7A and 7B, the sample tray 740, asmentioned above, includes a slit 780 defined within and extendingthrough the outwardly-bowed top portion 747 of the sample tray 740 forintroduction of the liquid sample into the interior pouch of theapparatus 700. Although the slit 780 extends through the sample tray 740(to provide access to the interior pouch of the apparatus 700), the slit780 does not penetrate or extend into the lid member 710 such that theslit 780 is the only access point for introducing the liquid sample intothe interior pouch of the apparatus 700. The slit 780 may be formed vialaser-cutting or in any other suitable fashion.

In use, the liquid sample, e.g., a 100 mL liquid sample including thetesting medium, is poured into the interior pouch of the apparatus 700through the slit 780. In order to introduce the liquid sample, theapparatus 700 is squeezed inwardly from the opposite side edges 702, 704thereof adjacent the outwardly-bowed top portion 747 to flex, or bendthe sample tray 740 (as indicated by arrows “A”) to define an arcuateconfiguration. In this configuration, the slit 780 is enlarged and aspace is established between the sample tray 740 and the lid member 710to enlarge the interior pouch opening defined by the slit 780.Alternatively, while grasping the apparatus 700, the outwardly-bowed topportion 747 of the sample tray 740 may be manipulated, e.g., bent,relative to the remainder of the sample tray 740 to likewise enlarge theinterior pouch opening defined via the slit 780.

With the liquid sample disposed within the interior pouch of theapparatus 700, the apparatus 700 may be fed into a heat sealer whichurges the lid member 710 into contact with the sample tray 740 such thatthe liquid sample is first evenly distributed into the small wells 744at full capacity, and such that the remainder of the liquid sample isevenly distributed into the large wells 748. The large wells 748 areonly partially filled, however, with the remaining portions of the largewells 748 occupied by air. Simultaneously with or near-simultaneouslywith the urging of the lid member 710 and the sample tray 740 intocontact with one another to distribute the liquid sample, heat appliedto the apparatus 700 via the heat sealer effects heat-sealing of the lidmember 710 to the sample tray 740 to sealingly enclose each of the wells744, 748. Incubation, result determination, and analysis may beperformed similarly as detailed above.

Referring to FIG. 8, multi-well sample testing apparatus 800 is similarto the apparatus 700 (FIGS. 7A and 7B) and generally includes a lidmember 810 and a sample tray 840. However, in contrast to the apparatus700 (FIGS. 7A and 7B), the sample tray 840 of the apparatus 800 includesa pair of reinforcement ribs 890 extending longitudinally on either sideof the small wells 844. Room for accommodating the reinforcement ribs890 may be provided by removing the outer-most rows of small wells 844on either side of the matrix, thus forming a 9×8 matrix of the smallwells 844. However, other suitable configurations, with or withoutremoving rows of wells, are also contemplated. The reinforcement ribs890 may be formed integrally with the sample tray 840, e.g., providingan increased thickness to form the reinforcement ribs 890.Alternatively, the reinforcement ribs 890 may be formed from anysuitable material, e.g., stainless steel, polymeric materials, etc., andmay be embedded within the sample tray 840, disposed on the interior orexterior-facing surface of the sample tray 840, disposed on the interioror exterior-facing surface of the lid member 810, or secured to theapparatus 800 in any other suitable fashion. The reinforcement ribs 890provide structural support to the apparatus 800 and inhibit twisting andother such manipulation of the apparatus 800. The reinforcement ribs 890also help maintain proper positioning of the apparatus 800 duringinsertion into and use of the heat sealer to seal the apparatus 800,thus helping to ensure an effective seal. The configuration and use ofthe apparatus 800 is otherwise similar to that of the apparatus 700(FIGS. 7A and 7B).

Referring to FIG. 9, multi-well sample testing apparatus 900 is similarto the apparatus 700, 800 (FIGS. 7A-7B and 8, respectively) andgenerally includes a lid member 910 and a sample tray 940. The apparatus900 differs from the apparatus 800 (FIG. 8) in the reinforcementthereof. More specifically, rather than providing the reinforcement ribs890 as in the apparatus 800 (see FIG. 8), the apparatus 900 includes anelongated, generally U-shaped reinforcement member 990 extending aboutthree perimeter edges of the sample tray 940, e.g., the first and secondside edges 902, 904, respectively, and the bottom edge 906. Thereinforcement member 990 may be formed integrally with the sample tray940, e.g., providing an increased thickness to form the reinforcementmember 990. Alternatively, the reinforcement member 990 may be formed asa wire or cable made from any suitable material, e.g., stainless steel,polymeric materials, etc., and may be embedded within the sample tray940, disposed on the interior or exterior-facing surface of the sampletray 940, disposed on the interior or exterior-facing surface of the lidmember 910, or secured to the apparatus 900 in any other suitablefashion. The reinforcement member 990 provides structural support to theapparatus 900 and inhibits twisting and other such manipulation of theapparatus 900. Further, the reinforcement member 990 helps maintainproper positioning of the apparatus 900 during insertion into and use ofthe heat sealer to seal the apparatus 900, thus helping to ensure aneffective seal. The configuration and use of the apparatus 900 isotherwise similar to that of the apparatus 700 (FIGS. 7A and 7B).

Turning to FIGS. 10A-10C, multi-well sample testing apparatus 1000 issimilar to the apparatus 700 (FIGS. 7A and 7B) and generally includes alid member 1010 and a sample tray 1040, which may include any of thefeatures of any of the other embodiments detailed herein, and/orcombinations thereof. The lid member 1010 and sample tray 1040 of theapparatus 1000 define generally rectangular configurations of similardimensions, except that the lid member 1010 and sample tray 1040 bothdefine outwardly-bowed top portions 1017, 1047, respectively.

The lid member 1010 of the apparatus 1000, more specifically, is formedfrom a plurality of layers: an adhesive layer 1012, a breathable filmlayer 1014, and a release liner 1018. The adhesive and breathable filmlayers 1012, 1014, respectively, may be configured similarly to thosedetailed above with respect to the lid member 110 of the apparatus 100(FIG. 1A). Exemplary embodiments of release liners will be described ingreater detail below with reference to FIGS. 11 and 12. The releaseliner 1018 is secured to the remainder of the lid member 1010 in anysuitable manner that permits the release liner 1018 to be peeled off ofor otherwise removed from the remainder of the lid member 1010, e.g.,the release liner 1018 may be secured to the remainder of the lid member1010 via casting, laminating, adhering, etc.

The apparatus 1000 defines a perforation 1011 that extends through theoutwardly-bowed top portion 1047 of the sample tray 1040 and partiallythrough the outwardly-bowed top portion 1017 of the lid member 1010.More specifically, the perforation 1011 extends through the adhesive andbreathable film layers 1012, 1014 of the lid member 1010, but not intothe release liner 1018. The perforation 1011 defines a wedge-shapeconfiguration that is wider adjacent the sample tray 1040 andprogressively decreases in width as the perforation 1011 extends furtherthrough the sample tray 1040 and into the adhesive and breathable filmlayers 1012, 1014 of the lid member 1010, although other configurationsare also contemplated. The perforation 1011 may be formed viakiss-cutting, or other suitable process.

Despite the perforation 1011 extending through the outwardly-bowed topportion 1047 of the sample tray 1040, the sample tray 1040 includes oneor more connectors 1043 extending between and interconnecting theportions 1042 a, 1042 b of the sample tray 1040 that are separated viathe perforation 1011. Connectors 1043 are formed integrally with theportions 1042 a, 1042 b of the sample tray 1040 and retain the portions1042 a, 1042 b of the sample tray 1040 in a generally planarconfiguration. As detailed below, the connectors 1043 are capable ofbeing snapped, or broken to permit removal of the portion 1042 a of thesample tray 1040 and a portion 1013 of the lid member 1010, therebyfacilitating removal of the release liner 1018 from the apparatus 1000.Further, the portion of sample tray 1040 adjacent perforation 1011(including connectors 1043) may define a rigid or more-rigidconfiguration to facilitate snapping, or breaking of connectors 1043. Asshown in FIG. 10A, two connectors 1043, one disposed towards each sideof the apparatus 1000, are provided, although other configurations arealso contemplated.

Similarly as detailed above with respect the apparatus 700 (FIGS. 7A and7B), the lid member 1010 and sample tray 1040 of the apparatus 1000 aresealingly engaged to one another along all four outer peripheral edgesof the apparatus 1000. However, rather than providing a separate slit,the portion of the perforation 1011 extending between the connectors1043 serves as the selectively enlargeable slit that permitsintroduction of the liquid sample into the interior of the apparatus1000.

In use, to introduce a liquid sample into the interior pouch defined bythe apparatus 1000, similarly as detailed above, the apparatus 1000 issqueezed inwardly from the opposite side edges thereof adjacent theperforation 1011 to flex, or bend the sample tray 1040 and enlarge thespacing between the sample tray 1040 and lid member 1010 adjacent theperforation 1011. As a result, the access opening to the interior pouchdefined between the sample tray 1040 and lid member 1010 is enlarged,facilitating introduction of the liquid sample into the interior of theapparatus 1000.

Once the liquid sample has been introduced into the apparatus 1000, theapparatus 1000 may be sealed using a heat sealer. During heat sealing,the release liner 1018 serves as an insulator to limit the amount ofheat conducted to the remainder of the lid member 1010 and/or otherportions of the apparatus 1000, thereby protecting the apparatus 1000from heat-damage and ensuring formation of an effective seal.Thereafter, the release liner 1018 is removed and the apparatus 100 isincubated. Alternatively, the release liner 1018 may be removed prior toheat sealing.

In order to remove the release liner 1018, the user grasps the body ofthe apparatus 1000 with one hand, and the outwardly-bowed top portions1047, 1017 of the sample tray 1040 and the lid member 1010 with theother hand, and bends the outwardly-bowed-top portions 1047, 1017 backonto the body portion of the apparatus 1000. The perforation 1011 servesas the hinge point for the bending of the outwardly-bowed-top portions1047, 1017 relative to the body portion of the apparatus 1000 and, uponsufficient bending, connectors 1043, which extend across the perforation1011, are snapped or broken, thereby separating the portions 1042 a,1042 b of the sample tray 1040 from one another. The snapping orbreaking of connectors 1043 also decouples the portion 1013 of theadhesive and breathable film layers 1012, 1014 of the lid member 1010from the remainder, e.g., the body, of the lid member 1010. However, asthe release liner 1018 is not interrupted by the perforation 1011, therelease liner 1018 remains intact.

With the portions 1042 a, 1013 of the sample tray 1040 and lid member1010, respectively, separated from the portion 1042 b and body portionof the lid member, respectively (except for the release liner 1018), theportions 1042 a, 1013 may be further pulled back towards the oppositeend of the apparatus 1000 to peel the release liner 1018 off of theremaining, e.g., the body, of the lid member 1010. Once the releaseliner 1018 (along with the portions 1042 b and 1013) have beenpeeled-off and removed, incubation, result determination, and analysismay be performed similarly as detailed above.

Turning now to FIG. 11, an embodiment of a lid member 1110 incorporatinga release liner 1118 and configured for use with the apparatus 1000(FIGS. 10A-10C), or any other suitable apparatus, is shown. The lidmember 1110 includes an adhesive layer 1112, a breathable film layer1114, and a release liner 1118. The adhesive layer 1112 and breathablefilm layer 1114 may be configured similarly as any of the above-detailedembodiments, and may be joined via co-extrusion, or other suitableprocess. The release liner 1118 includes a relatively thin siliconelayer 1119 a, and a relatively thick clay-coated paper layer 1119 bdisposed on the silicone layer 1119 a. The silicone layer 1119 a allowsfor the initial retention of the release liner 1118 about the adhesiveand breathable film layers 1112, 1114, but also facilitates the removaltherefrom upon peeling of the release liner 1118. The breathable filmlayer 1114 is extruded directly onto the silicone layer 1119 a of therelease liner 1118 and the adhesive layer 1112 is extruded directly ontothe breathable film layer 1114. Alternatively, the breathable film layer1114 and the adhesive layer may be co-extruded directly onto the releaseliner 1118.

FIG. 12 illustrates another embodiment of a lid member 1210incorporating a release liner 1218 and configured for use with theapparatus 1000 (FIGS. 10A-10C), or any other suitable apparatus. The lidmember 1210 includes an adhesive layer 1212, a breathable film layer1214, and a release liner 1218. The adhesive layer 1212 and breathablefilm layer 1214 may be configured similarly as any of the above-detailedembodiments, and may be joined via co-extrusion, or other suitableprocess. The release liner 1218 includes a relatively thin siliconelayer 1219 a, a relatively thick clay-coated paper layer 1219 b, and apolyester layer 1219 c having a thickness less than that of theclay-coated paper layer 1219 b but greater than that of the siliconelayer 1219 a. The breathable film layer 1214 is extruded directly ontothe silicone layer 1219 a of the release liner 1218 and the adhesivelayer 1212 is extruded directly onto the breathable film layer 1214.Alternatively, the breathable film layer 1214 and the adhesive layer1212 may be co-extruded directly onto the release liner 1218. Further,the silicone, clay-coated paper, and polyester layers 1219 a, 1219 b,1219 c may be coupled to one another via laminating, or other suitableprocess.

Referring generally to FIGS. 13A-16D, and as mentioned above, thevarious apparatus detailed herein are configured for use with a heatsealer to seal a portion of the liquid sample within each of the wellsof the apparatus. In order to facilitate the heat sealing of theapparatus, a receptacle configured to receive the apparatus may beutilized. The receptacle maintains the apparatus in a desiredorientation, inhibits bending or twisting of the apparatus, and, inconjunction with the heat sealer, guides the apparatus through the heatsealer to ensure formation of an effective seal without damaging theapparatus. Various embodiments of such receptacles are detailed below.Each of these receptacles includes a base portion and a releasablyengagable cover flap.

In embodiments where the apparatus to be sealed defines a relativelythin configuration, is more prone to sticking or misalignment, and/orincludes a more heat-sensitive lid member, e.g., in embodiments where arelease liner is not provided, the cover flap is utilized as a protectorthat protects the lid member of the apparatus during heat sealing.However, in other embodiments where such concerns are minimal, e.g.,where a release liner is provided or the apparatus defines a more rigidor robust configuration, the base portion of the receptacle may beutilized without the cover flap.

Referring to FIGS. 13A-13C, a receptacle 1300 is shown configured foruse with an apparatus 1301. The apparatus 1301 is shown generically, asit is envisioned that any suitable apparatus, such as any of thosedetailed above, may be utilized in conjunction with the receptacle 1300to facilitate sealing of the apparatus 1301 when put through a heatsealer. The receptacle 1300 includes a base portion 1320 and a coverflap 1340 that is releasably engagable with the base portion 1320. Thebase portion 1320 is configured to receive the apparatus 1301 therein,while the cover flap 1340 is configured to sit atop the lid member 1302of the apparatus 1301 to enclose the apparatus 1301 within thereceptacle 1300 and protect the lid member 1302 of the apparatus 1301during heat sealing.

The base portion 1320 of the receptacle 1300 defines a generallyrectangular configuration having an upper surface 1322 and a cavity 1324recessed within the upper surface 1322. The cavity 1324 is dimensionedsimilarly to the apparatus 1301 and is configured to receive theapparatus 1301 therein. In some configurations, the cavity 1324 isdimensioned such that the outer peripheral edge of the apparatus 1301serves as a lip that is seated on the upper surface 1322 of the baseportion 1320 while the remainder of the apparatus 1301 is seated withinthe cavity 1324. The cavity 1324 may further include dividers, alignmentstructures, etc. (not explicitly shown) disposed therein that areconfigured for positioning between the wells of the apparatus 1301 tohelp ensure proper placement and alignment of the apparatus 1301 withinthe base portion 1320 of the receptacle 1300. The base portion 1320further includes an engagement slot 1326 disposed towards an end thereofand extending transversely along a portion of the end of the baseportion 1320. The engagement slot 1326 defines a shoulder 1328configured to pivotably engage the cover flap 1340, as detailed below.

The cover flap 1340, shown in FIG. 13B, includes a generally planar bodyportion 1342, an engagement pin 1344, and a flange portion 1346extending between and engaging the body portion 1342 and the engagementpin 1344 to one another. The body portion 1342 and flange portion 1346,which is integral with the body portion 1342, may be formed from asilicone-based material, or any other suitable material(s). The baseportion 1320 (FIG. 13A) may likewise be formed from a silicone-basedmaterial or other suitable material(s) similar to or different from thatof the cover flap 1340. The engagement pin 1344 may be formed fromstainless steel or other suitable material suitable for use in a heatsealer. Further, the flange portion 1346 may be secured to theengagement pin 1344 via an adhesive, may be looped about the engagementpin 1344, or may be secured thereto in any other suitable fashion.

Referring to FIG. 13C, in order to engage the cover flap 1340 with thebase portion 1320, the engagement pin 1344 of the cover flap 1340 isinserted into the engagement slot 1326 of the base portion 1320 undersufficient urging to expand the engagement slot 1326 and permit passageof the engagement pin 1344 beyond the shoulder 1328, such that theengagement pin 1344 is snapped into engagement within theenlarged-diameter portion 1329 of the engagement slot 1326 adjacent theshoulder 1328. In this position, the shoulder 1328 pivotably retains theengagement pin 1344 therein, while permitting the cover flap 1340 topivot from an open position (FIG. 13A) for insertion and removal of theapparatus 1301 (FIG. 13A) from the base portion 1320, and a closedposition, wherein the cover flap 1340 is disposed atop the lid member1302 of the apparatus 1301 (FIG. 13A) to enclose the apparatus 1301(FIG. 13A) within the receptacle 1300.

Turning to FIG. 14, another embodiment of a receptacle 1400 provided inaccordance with the present disclosure is shown. The receptacle 1400includes a base portion 1420 and a cover flap 1440 and is similar to thereceptacle 1300 (FIGS. 13A-13C) except for the engagement mechanism forcoupling the base portion 1420 and cover flap 1440 to one another. Forpurposes of brevity, only such differences are detailed below.

The base portion 1420 of the receptacle 1400 includes a pair of fingers1422, 1424 extending from an end thereof, one positioned towards eachend corner of the base portion 1420. The fingers 1422, 1424 arespaced-apart from the base portion 1420 and extend towards one anotherto define an engagement area 1423, 1425 between the respective finger1422, 1424 and the base portion 1420.

The cover flap 1440 includes a generally planar body portion 1442 and anengagement pin 1444 coupled to the body portion 1442, e.g., via anintermediate flange portion or directly thereto. The ends of theengagement pin 1444 are configured for receipt within the engagementareas 1423, 1425 of the base portion 1420 in pivotable, snap-fitengagement therewith to couple the cover flap 1440 to the base portion1420.

FIG. 15 illustrates another embodiment of a receptacle 1500 provided inaccordance with the present disclosure. The receptacle 1500 includes abase portion 1520 and a cover flap 1540 and is similar to the receptacle1300 (FIGS. 13A-13C) except for the engagement mechanism for couplingthe base portion 1520 and cover flap 1540 to one another. For purposesof brevity, only such differences are detailed below.

The base portion 1520 of the receptacle 1500 includes a slot 1522defined therein towards and end thereof that extends transversely acrossthe base portion 1520. The slot 1522 includes a relatively narrow neckportion 1524 extending adjacent the mouth of the slot 1522, and acylindrical, relatively larger diameter engagement portion 1526extending along the nadir of the slot 1522.

The cover flap 1540 includes a generally planar body portion 1542 and anengagement pin 1544 coupled to the body portion 1542, e.g., via anintermediate flange portion or directly thereto. The engagement pin 1544defines a generally cylindrical configuration that approximates thediameter of the cylindrical engagement portion 1526 of the slot 1522defined within the base portion 1520 but is larger than the width of theneck portion 1524.

In order to engage the cover flap 1540 with the base portion 1520, theengagement pin 1544 of the cover flap 1540 is inserted into the slot1522 of the base portion 1520 under sufficient urging to expand the neckportion 1524 and permit passage therethrough, ultimately such that theengagement pin 1544 is seated within the cylindrical portion 1526 of theslot 1522, thereby pivotably coupling the cover flap 1540 to the baseportion 1520.

Referring to FIGS. 16A-16D, another embodiment of a receptacle 1600provided in accordance with the present disclosure is shown. Thereceptacle 1600 includes a base portion 1620 and a cover flap 1640 andis similar to the receptacle 1300 (FIGS. 13A-13C) except for theengagement mechanism for coupling the base portion 1620 and cover flap1640 to one another. For purposes of brevity, only such differences aredetailed below.

The base portion 1620 of the receptacle 1600 defines a cavity having aplurality of discrete chambers 1621 a arranged to define a configurationcomplementary to that of the apparatus 1601 configured for receipttherein such that each of the chambers 1621 a receives one of the wellsof the apparatus 1601. The base portion 1620 also defines a cut-out 1621b at a first end thereof that is configured to facilitate the insertionand removal of the apparatus 1601 therefrom. Although not shown, eitheror both of these features may be provided for use with theabove-detailed receptacles.

The base portion 1620 of the receptacle 1600 further includes aplurality of spaced-apart fingers, e.g., three, spaced-apart fingers1622, 1624, 1626, extending from the second end (the opposite end as thecut-out 1621 b) thereof. As illustrated, a first finger 1622 is disposedtowards one side of the base portion 1620, a second finger 1624 isdisposed towards the other, opposite side of the base portion 1620, anda third finger 1626 is disposed intermediate the first and secondfingers 1622, 1624, although other configurations, including greater orfewer fingers, are also contemplated. As the fingers 1622, 1624, 1626are spaced-apart relative to one another, first and second bays 1628,1629 are defined between the fingers 1622, 1626 and the fingers 1624,1626, respectively. Further, each of the fingers 1622, 1624, 1626defines a lumen 1623, 1625, 1627 extending transversely therethrough.The lumens 1623, 1625, 1627 are aligned with one another in co-axialrelation.

The cover flap 1640 includes a generally planar body portion 1642 and apair of spaced-apart flanges 1646 extending from an end of the bodyportion 1642. Each of the flanges 1646 is formed integral with the bodyportion 1642 and includes a tubular end portion 1647 defining a lumen1648 extending therethrough. The lumens 1648 are aligned in co-axialrelation relative to one another.

In order to engage the cover flap 1640 with the base portion 1620, thecover flap 1640 is positioned such that the flanges 1646 thereof aredisposed within the first and second bays 1628, 1629 and such that thelumens 1648 are aligned in co-axial relation with the lumens 1623, 1625,1627. Thereafter, an engagement pin 1644 is inserted through the lumen1623, one of the lumens 1648, the lumen 1627, the other of the lumens1648, and the lumen 1625 (although the reverse order of insertion isalso contemplated) to pivotably couple the base portion 1620 and coverflap 1640 to one another.

From the foregoing and with reference to the various figure drawings,those skilled in the art will appreciate that certain modifications canalso be made to the present disclosure without departing from the scopeof the same. While several embodiments of the disclosure have been shownin the drawings, it is not intended that the disclosure be limitedthereto, as it is intended that the disclosure be as broad in scope asthe art will allow and that the specification be read likewise. Forexample, while the lid member is shown in the illustrated embodimentsincluding a backing layer that is preferably paper, embodiments wherethe backing layer is not provided are also contemplated, as is formingthe backing layer from alternate materials such as woven and non-wovenfabrics. Therefore, the above description should not be construed aslimiting, but merely as exemplifications of particular embodiments.Those skilled in the art will envision other modifications within thescope and spirit of the claims appended hereto.

What is claimed is:
 1. A sample testing apparatus, comprising: a sample tray defining a planar surface and a plurality of wells recessed relative to the planar surface; and a lid member, including: an adhesive layer configured to be sealed to the planar surface of the sample tray; a breathable film layer disposed about the adhesive layer; and a backing layer disposed about the breathable film layer.
 2. The apparatus according to claim 1, wherein the sample tray is formed from a permeable, transparent, non-toxic material.
 3. The apparatus according to claim 2, wherein the sample tray is formed from a blend of a styrene butadiene copolymer and general purpose polystyrene.
 4. The apparatus according to claim 1, wherein the adhesive layer is formed from a material that is permeable and capable of being heat sealed to the planar surface of the sample tray.
 5. The apparatus according to claim 4, wherein the adhesive layer is formed from a material that is transparent.
 6. The apparatus according to claim 4, wherein the adhesive layer is formed from ethylene vinyl acetate or modified ethylene vinyl acetate.
 7. The apparatus according to claim 1, wherein the adhesive layer includes a plurality of perforations, each perforation corresponding to and aligned with one of the wells defined within the sample tray.
 8. The apparatus according to claim 1, wherein the breathable film layer is formed from a material that is permeable.
 9. The apparatus according to claim 8, wherein the breathable film layer is formed from a material that is transparent.
 10. The apparatus according to claim 8, wherein the breathable film layer is formed from a thermoplastic copolyester based elastomer.
 11. The apparatus according to claim 1, wherein the backing layer is a paper backing layer including a paper that is permeable, insulating, and capable of adhesive-less bonding to the breathable film layer.
 12. The apparatus according to claim 11, wherein the backing layer includes one of a kraft paper, a clay-coated paper, or an offset paper.
 13. The apparatus according to claim 1, wherein the backing layer is configured as a release liner that is removable from the breathable film layer.
 14. The apparatus according to claim 13, wherein the release liner includes a paper layer and a silicone layer.
 15. The apparatus according to claim 14, wherein the release liner further includes a polyester layer.
 16. The apparatus according to claim 13, wherein a perforation extending through the sample tray and partially through the lid member is provided to facilitate removal of the release liner.
 17. The apparatus according to claim 16, wherein at least one connector interconnects portions of the sample tray on either side of the perforation.
 18. The apparatus according to claim 17, wherein the at least one connector is breakable to permit removal of the release liner.
 19. The apparatus according to claim 1, wherein the plurality of wells include at least a first set of wells and a second set of wells, the first and second sets of wells defining different configurations.
 20. The apparatus according to claim 1, wherein the lid member is initially sealed to the sample tray about a first peripheral side, a second peripheral side, and a bottom end of the sample tray to define a pouch, and wherein an open top end of the sample tray is configured to permit introduction of a liquid sample into the pouch.
 21. The apparatus according to claim 20, wherein the lid member is sealed about the sample tray by heat sealing the adhesive layer to the planar surface, thereby sealing each of the wells with a portion of the liquid sample therein.
 22. The apparatus according to claim 1, wherein the lid member is initially sealed to the sample tray entirely about an outer perimeter of the planar surface of the sample tray to define a pouch therebetween, and wherein the sample tray defines a slit extending therethrough that is configured to permit introduction of a liquid sample into the pouch.
 23. The apparatus according to claim 1, further including at least one reinforcement member coupled to the sample tray, the at least one reinforcement member configured to provide structural support to the sample tray.
 24. The apparatus according to claim 1, wherein the sample tray defines at least one bridge fluidly connecting adjacent wells to facilitate the capture of an air bubble within at least one of the adjacent wells upon sealing of the lid member with the sample tray.
 25. A method of testing a sample, comprising: providing a sample testing apparatus, including: a sample tray defining a plurality of wells including at least a first set of wells having a first configuration and a second set of wells having a second configuration, wherein the first and second configurations are different from one another; and a lid member sealed to the sample tray about both sides and at least one end of the sample tray to define a pouch; introducing a predetermined volume of a liquid sample into the pouch of the sample testing apparatus; and sealing the lid member to the sample tray to seal each of the plurality of wells such that each of the wells of the first set of wells is filled to capacity with a first portion of the liquid sample and such that a reminder of the liquid sample is evenly distributed into the wells of the second set of wells, wherein a volume of liquid sample in each of the wells of the second set of wells is less than a volume capacity of each of the wells in the second set of wells.
 26. The method according to claim 25, wherein sealing the lid member to the sample tray is performed via heat sealing.
 27. The method according to claim 26, further including positioning the sample tray in a receptacle prior to heat sealing.
 28. The method according to claim 27, further including enclosing the sample tray within the receptacle using a cover flap of the sample tray.
 29. The method according to claim 25, further comprising incubating the sealed sample testing apparatus.
 30. The method according to claim 25, further comprising peeling off a backing layer of the lid member prior to incubating the sealed sample testing apparatus.
 31. The method according to claim 30, wherein peeling off the backing layer includes grasping a perforated portion of the sample testing apparatus and moving the perforated portion relative to the sample testing apparatus to peel off the backing layer.
 32. The method according to claim 30, further comprising counting a number of positive wells and determining a result based upon the number of positive wells.
 33. The method according to claim 25, wherein the lid member is sealed to a bottom of the sample tray and unsealed at a top of the sample tray, and wherein introducing the predetermined volume of the liquid sample into the pouch includes introducing the predetermined volume through the unsealed top.
 34. The method according to claim 25, wherein the lid member is sealed about a perimeter edge of the sample tray, and wherein introducing the predetermined volume of the liquid sample into the pouch includes introducing the predetermined volume through a slit defined within the sample tray. 